Kiryushkin Alexey S, Ilina Elena L, Guseva Elizaveta D, Pawlowski Katharina, Demchenko Kirill N
Laboratory of Cellular and Molecular Mechanisms of Plant Development, Komarov Botanical Institute, Russian Academy of Sciences, 197376 Saint Petersburg, Russia.
Department of Ecology, Environment and Plant Sciences, Stockholm University, 10691 Stockholm, Sweden.
Plants (Basel). 2021 Dec 24;11(1):51. doi: 10.3390/plants11010051.
CRISPR/Cas-mediated genome editing is a powerful tool of plant functional genomics. Hairy root transformation is a rapid and convenient approach for obtaining transgenic roots. When combined, these techniques represent a fast and effective means of studying gene function. In this review, we outline the current state of the art reached by the combination of these approaches over seven years. Additionally, we discuss the origins of different strains that are widely used for hairy root transformation; the components of CRISPR/Cas vectors, such as the promoters that drive Cas or gRNA expression, the types of Cas nuclease, and selectable and screenable markers; and the application of CRISPR/Cas genome editing in hairy roots. The modification of the already known vector pKSE401 with the addition of the rice translational enhancer and the gene encoding the fluorescent protein DsRed1 is also described.
CRISPR/Cas介导的基因组编辑是植物功能基因组学的一种强大工具。毛状根转化是获得转基因根的一种快速便捷的方法。当这两种技术结合使用时，它们代表了一种研究基因功能的快速有效的手段。在本综述中，我们概述了这两种方法结合七年来所达到的当前技术水平。此外，我们还讨论了广泛用于毛状根转化的不同菌株的来源；CRISPR/Cas载体的组成部分，如驱动Cas或gRNA表达的启动子、Cas核酸酶的类型以及选择和筛选标记；以及CRISPR/Cas基因组编辑在毛状根中的应用。还描述了通过添加水稻翻译增强子和编码荧光蛋白DsRed1的基因对已知载体pKSE401进行的修饰。
